System for growing fungal materials

ABSTRACT

A system for growing fungi, the system comprising a nutritive vehicle, a porous material, an administrable space, fungal tissue comprising fungal hyphae having a growth pattern, the fungal tissue connecting said nutritive vehicle to said porous material to said administrable space, wherein the fungal tissue within said space defines at least one successive fungal material layer; and a chemically or physically altered separated portion of fungal material, the separated portion separated from said fungal tissue.

RELATED APPLICATIONS

This application is a Continuation application based on U.S.Nonprovisional application Ser. No. 17/326,742, filed May 21, 2021, andgranted as U.S. patent Ser. No. 11/310,968 on Apr. 26, 2022, and whichis a Continuation application based on U.S. Nonprovisional applicationSer. No. 17/081,745, filed Oct. 27, 2020, and granted as U.S. Pat. No.11,013,189 on May 25, 2021, and which is a Continuation applicationbased on U.S. Nonprovisional application Ser. No. 15/884,788, filed Jan.31, 2018 and granted as U.S. Pat. No. 10,842,089 on Nov. 24, 2020, whichwas a Continuation application of U.S. Nonprovisional application Ser.No. 15/650,779, filed Jul. 14, 2017 and granted Jun. 23, 2020 as U.S.Pat. No. 10,687,482, which claims priority to U.S. Provisional PatentApplication 62/362,462, filed Jul. 14, 2017, the disclosures of whichare incorporated herein by reference in their entirety.

BACKGROUND

The present disclosure relates generally to methods for growing fungalmaterials and objects therefrom, and in particular to a method forproducing fungal materials and objects with variable shape, size,thickness, density, flexibility, and other predetermined qualitiesthrough directed and controlled tissue development and post-growthprocessing.

The material of a plant's body is formed from carbon and other elementsfiltered from the air, then bound together into sugars and other largemolecules using energy from the sun. Mushrooms lack the ability tosynthesize their bodies directly from sunlight and, like animals, needto consume things that were once alive in order to survive and grow.Filamentous fungi grow their bodies as an expanding and interconnectedweb of threadlike cells (called hyphae) directly within the food theyare in the process of consuming as nutrients. The threadlike hyphae ofthe growing fungi exude strong enzymes and other agents into the wood orother substrate material it is living within and dissolves the molecularbonds that provide these substrates their structure. The fungus absorbsdissolved nutrients taken from the cellulose, lignin, and othersubstances present, which it then uses to build chitin, the resilientand strong protein that comprises its own hyphal walls. Chitin, likecellulose and keratin, is a naturally forming polymer that is found inthe toughest organic tissues. In addition to being found in all fungi,chitin helps create the durable and flexible exoskeletons of insects andshellfish. These remarkable qualities are due in great part to thestructure of the tissues' constituent parts as well as the properties ofthe materials that make them up.

Filamentous fungi have the natural tendency to join together smallerpieces of branching, colonial hyphae into a larger constituent whole,assembling and weaving strands and sheets of tissues called mycelium.Mycelium can adhere to, and possibly engulf, any other materials itcomes in contact with through the extension of hyphae that use neighborsensing and searching functions as guidance in their exploration intospace beyond sources of nutritional sustenance. Like cement and plaster,fungal tissue will bind, harden and set into a variety of solidifiedconfigurations through the natural biological functions of mycelialgrowth and self-adhesion.

Fungal tissue can quickly be amplified to an enormous volume if providedwith the appropriate living conditions. These conditions include thenutrients that might be available to the organism, the possible gasgradients within the growth environment and the humidity, light, andtemperatures the organism might be exposed to as it takes form. Fungiare very sensitive to their surroundings, and by altering subtle factorsit is possible to prompt their tissue to express a range of variablydetermined physical characteristics.

Fungi are very sensitive to chemicals present in their environment, andhave the ability to alter the directions and vigor of growth ofexpanding hyphae as demonstrated through chemotaxic avoidance orattraction. Fungi are also very sensitive to other stimuli in theirenvironment, and have the ability to alter directions and vigor ofgrowth of expanding hyphae in response to gravitropic, thermotropic,thigmotropic, phototropic, and hydrotropic stimuli.

A substrate colonized with fungal hyphae, if provided adequate enclosureand environmental controls, will in a matter of one to three daysgenerate a layer of fungal hyphae growing from the top of said substratethat will expand into space as a layer in a fuzzy and undifferentiatedmanner. This undifferentiated layer of hyphae, if left to continuegrowing, will soon advance in development and differentiate intospecialized tissues determined to become fruit bodies or othersporocarp-producing structures.

Fungus-based materials and composites can be propagated on readilyavailable agricultural waste, using principles and techniques that arewell established with regard to growing filamentous fungi for humanconsumption and industry.

DESCRIPTION OF THE RELATED ART

Current forms of manufacturing of polymeric materials including animalskins and vinyl create environmental problems in manufacturing andrecycling or disposal at the end of a material's utility. There is aneed for new means of creating large volumes of polymeric materials thatcan be produced in ways that use fewer resources and otherwise diminishthe environmental impacts of current practices.

Composite materials may be formed from fungus by mixing an inoculumincluding a preselected fungus with discrete particles and a nutrientmaterial capable of being digested by the fungus. Furthermore, fungalprimordium may be enclosed and grown in a mold to obtain a mass offungal tissue in the form of low density chitinous material. Methodsalso exist for limiting the advancement of development of fungal tissuestowards fruit bodies and sporocarp supporting structures through theadministration of carbon dioxide/oxygen gradients and relativetemperatures in which the organism is growing.

Unlike fungal tissue composites which are composed of lignocellulosicwaste materials, fungal cellular tissue, and other ingredients, thefungal tissue materials and composites described herein can consistalmost entirely of fungal tissues. This fungal material can be used inproducts that are currently made with ethylene vinyl acetate foams,polyvinyl chloride plastics, polyurethane foams, amongst others.

The state of the art in creating materials that are composed of fungaltissues in their manufacture is limited by the capacity of liquid orsubmerged cultures in the propagation of filamentous hyphae, which arethen harvested and cultivated on secondary scaffolds and incubators. Themethod described herein can easily be adapted to the leading globalmethod for producing fungal mycelium and edible mushrooms: solid statefermentation. Solid lignocellulosic substrates are easy to obtain,relatively inexpensive, and can be processed in extremely large volumes.Using solid-state nutrients allows for the making of 3-D molds anddirectly growing 3-D forms into pressed or otherwise structured andmolded forms. This invention provides novel means of producing fungusbased materials at larger volumes in size and scale, with greaterdeterminations and controls for material function, using methodologiesand hardware that are simpler and a more efficient means of productionthan are currently available.

In recent years, numerous methods and systems have been used for thelarge-scale production of composites grown with mycelium. Conventionalmethods for the production of materials made from fungal materials arelimited:

Furthermore, conventional approaches do not produce a biologicallyactive fungal tissue material. Most methods for producing pure samplesof fungal tissue materials are reliant on submerged liquid cultures orrafts to cultivate significant volumes of hyphae, or require growinghyphae directly within a support matrix under very particular ecologicalcontrol regimens. Without the use of a semi-permeable intermediate layerof material, the growing mycelium has been shown to unevenly generate,bind, and fuse to the heterogeneously ordered fibers that constitute thesubstrate, leading to a disrupted surface of the substrate, and highlevels of heterogeneity and shear stresses in the growing sheet offungal tissue. An intermediate layer can be considered as a layerthrough which fungal hyphae might grow or be prohibited to do so.

Additionally, conventional methods do not allow for easy manipulation ofgrowing fungal hyphae to produce materials with desired qualities suchas lattices, and other two or three-dimensional engineered structures aspart of their constitution. Neither do conventional approaches accountfor post-growth processing of fungal tissue materials, without which theresulting outcome will exhibit poor mechanical characteristics when dry,particularly exhibiting low flexibility and high brittleness.

There is thus a need for a method for producing a fungal tissuematerials formed of variable thickness, density and other qualities foruse by various industrial processes and applications. Such a methodwould include an intermediate layer set upon a growing expression ofhyphae from the surface of a colonized substrate. Such a method wouldinclude an intermediate layer that controls the interaction of thefungal material's structure with the substrate, for example by providinga membrane permeable to the fungal hyphae but not to substrate particlesand fibers, thus allowing for easy separation of the two and eliminatingadditional processes for removing substrate particles and fibers fromthe harvested fungal materials. Such a method would include anintermediate layer that prevents the fungal material from permanentlyadhering to the substrate and which prevents damaging or tearing of thesubstrate when the fungal material is removed therefrom. Such a methodwould further allow the substrate to be reused for growing other fungalmaterials Such a method would further allow the manipulation of fungalmaterials to grow in particular and predetermined directions such aspurposefully engineered structures, lattices, and other two orthree-dimensional orientations of matter. Such a method would furtherallow the formation of multiple sheets of fungal material which can bestacked or arranged in a continuous form and finally, would be aninexpensive, simple, means of producing materials with lessenvironmental impact than conventional methods.

SUMMARY OF THE INVENTION

Certain embodiments of the present invention provide methods for growinga fungus polymer matrix that is comprised predominately of fungaltissues. The resultant material is a flexible and soft, high-densityamorphous polymer that can serve in applications that are currentlyserved by synthetic plastics and foams as well as animal skins. Thesubstrate utilized may comprise a mixture of discrete particles andnutrients and in some instances, is provided in an enclosure. A fungalinoculum may be prepared using a desired fungi strain. Hyphae from adefined species of filamentous fungi are propagated from a colonizablesubstrate that has been inoculated with said chosen fungi. Preferredspecies include the Ganodermas, the order Polyporales generally, andincluding all saprobic fungal candidates that derive sustenance fromlignin and cellulose-rich sources. The fungal inoculum may be introducedinto the substrate within the enclosure or prior to being introduced tothe enclosure so as to provide an even distribution of the fungusthroughout. The substrate is left to colonize. An intermediate layer isestablished on an open surface of the colonized substrate to control theinteraction of the forming fungal tissue structure with the substrate.The presence of a uniform intermediate material atop the substrateenables a consistent surface from which the fungal tissues may grow,supporting uniform expansion of the fungal hyphae into the environment,and providing a determined space for manipulation by chemical andphysical controls. Live fungal hyphae grow from the substrate andthrough the intermediate layer. In some instances, the living tissuesthat extend through the intermediate layer are manipulated to achieve amaterial having a desired thickness, shape, size and qualities.

The intermediate layer may be delaminated from the nutrient source outof which it has grown to terminate further growth of the material, orthe fungal tissue layer may be delaminated from the intermediate layer,which is left in place and optionally reused. The resultant livingfungal tissue structures may optionally be fused with other livingfungal tissue structures to create two-dimensional and three-dimensionalstructures. The final fungal tissue may then be subjected to post-growthprocessing to achieve desired properties for downstream usage.

The fungal substrate precursor material may be cultivated in eitherbatch or continuous processes and the fungal tissues may be modified anddirected during growth in order to achieve uniform characteristicsacross a surface, or be engineered to take on distinct local qualitiesthrough manipulation of growing tissue, or the addition of particles,fibers, meshes, fabrics, and other additives, armatures, and components.

Fungal tissue sheets may be processed via cutting or other formingmethods to obtain two-dimensional features and reliefs, or individualsheets may be stacked and grown together to form three-dimensionalfeatures, or composed with reinforcements or other structural amendmentsthat may be incorporated into a growing tissue.

It is a first objective of the present invention to provide a method forproducing a fungal materials and structures of variable shape,thickness, density, flexibility, and other predetermined qualities forindustrial applications. Such a method includes the production of bothtwo-dimensional and three-dimensional shapes by varying the shape of theenclosure, nutritive vehicle, intermediate layer and the conformation ofproduced fungal materials.

Another objective of the present invention is to provide an intermediatelayer on an exposed surface of a substrate to promote uniform growth offungal material.

Another objective of the invention is to provide an intermediate layerthat may be separable from the substrate with minimal force andprocesses required.

Another objective of the invention is to provide an intermediate layerfrom which the fungal material may be easily separable and which remainsattached to the substrate such that it may be re-used to grow additionalfungal material.

Yet another objective of the invention is to provide an intermediatelayer that prevents the fungal material from permanently adhering to thesubstrate and prevents damaging or tearing of the substrate whenremoving the fungal material.

Yet another objective of the invention is to provide a method thatdirects the growth of fungal material in particular and predeterminedpatterns such as orthogonal structures, lattices and other two orthree-dimensional structures.

Yet another objective of the invention is to provide a method thatallows for the fused growth of multiple fungal materials or structures,which can be stacked, folded, or otherwise arranged into 2D or 3D forms.

A yet another objective of the invention is to provide a method foradding composite materials to the fungal materials or structures toachieve desired material properties.

Yet another objective of the invention is to provide a method forprocessing the raw fungal material after growth to achieve desiredcharacteristics such as improved flexibility and tensile strength.

Yet another object of the invention is to provide methods for creatingan intermediate layer comprising a woven or porous surface that form apartial or complete laminate upon the surface of a colonized fungalsubstrate.

Yet another object of the invention is to provide a method fordelaminating a fungal material with ease and lack of damage to thesubstrate that fueled the growth of said fungal material.

Yet another object of the invention is to provide a method for dryingfungal materials (e.g. through using forced convection or conduction) todeactivate the fungus and prevent further growth.

Yet another object of the invention is to provide a method for the postprocessing of fungal materials to modify structure or chemicalcomposition, thereby conferring physical qualities according to theirdesired application.

Yet another object of this invention is to provide a fungal biopolymermaterial for use in functional products.

Yet another object of the invention is to provide simple, economicaltechniques for making fungal biopolymer products.

Yet another object of the invention is to provide a method for thegrowing of a fungus-based polymer matrix.

Yet another object of the invention is to provide a method for thegrowing of a fungus-based polymer matrix in the form of a composite.

Yet another object of the invention is to provide a material that canact as an analog to synthetic plastic materials, foams, and animalskins.

These and other aspects, objects, and features of the present inventionwill be understood and appreciated by those skilled in the art uponstudying the following specification, claims, and appended drawings.

The following detailed description together with accompanying figureswill provide a better understanding of the nature and advantages of thepresent invention.

DESCRIPTIONS OF THE FIGURES

In order to enhance clarity and improve understanding of the variouselements and embodiments of the invention, elements in the figures havenot necessarily been drawn to scale. Furthermore, elements that areknown to be common and well understood to those in the industry are notdepicted in order to provide a clear view of the various embodiments ofthe invention. Thus, the drawings are generalized in form in theinterest of clarity and concision.

FIG. 1 illustrates a block diagram of a method of producing fungalmaterial in accordance with the preferred embodiment of the presentinvention;

FIG. 2 illustrates a flowchart of the method of producing fungalmaterial in accordance with the preferred embodiment of the presentinvention;

FIG. 3 illustrates an enclosure for placing a nutritive vehicle inaccordance with the preferred embodiment of the present invention;

FIG. 4 illustrates the nutritive vehicle placed within the enclosure inaccordance with the preferred embodiment of the present invention;

FIG. 5A illustrates an intermediate layer being placed on an exposedsurface of the nutritive vehicle in accordance with the preferredembodiment of the present invention;

FIG. 5B illustrates the intermediate layer on an exposed surface of thenutritive vehicle in accordance with the preferred embodiment of thepresent invention;

FIG. 5C illustrates a composite material being placed onto fungalmaterial that has grown through the intermediate layer in accordancewith the preferred embodiment of the present invention;

FIG. 6A illustrates the composite material on top of the fungal materialand intermediate layer and an enclosure lid with a plurality of openingsin accordance with the preferred embodiment of the present invention;

FIG. 6B illustrates fungal tissue grown through the composite materialand the enclosure lid with a plurality of openings in accordance withthe preferred embodiment of the present invention;

FIG. 7 illustrates the enclosure closed with the lid in accordance withthe preferred embodiment of the present invention;

FIG. 8 illustrates a first dense network of hyphae growing off of thenutritive vehicle and through the intermediate layer in accordance withthe preferred embodiment of the present invention;

FIG. 9 illustrates the first dense network of hyphae shown in FIG. 8being rolled flat in a first direction on an exposed surface of theintermediate layer to form a first flattened fungal material inaccordance with the preferred embodiment of the present invention;

FIG. 10 illustrates a second dense network of hyphae regrown from thenutritive vehicle and through the intermediate layer and the firstflattened fungal material shown in FIG. 9 in accordance with thepreferred embodiment of the present invention;

FIG. 11 illustrates the second dense network of regrown hyphae afterbeing rolled in a second direction to form a second flattened fungalmaterial in accordance with the preferred embodiment of the presentinvention;

FIG. 12 illustrates a third dense network of hyphae regrown from thenutritive vehicle and through the intermediate layer and first andsecond flattened fungal materials shown in in FIG. 11 in accordance withthe preferred embodiment of the present invention;

FIG. 13A is a perspective view of the hyphae in a growing state;

FIG. 13B is a perspective view of the hyphae being flattened in a firstdirection;

FIG. 13C is a perspective view of hyphae in a growing state;

FIG. 13D is a perspective view of the hyphae being flattened in a seconddirection;

FIG. 13E is a perspective view of hyphae in a growing state;

FIG. 13F is a perspective view of the hyphae being flattened in a thirddirection;

FIG. 13G is a perspective view of the hyphae flattened in multipledirections;

FIG. 14A is a first cross-sectional illustration of the fungal materialabout to be pierced with a pointed element descending verticallytherethrough;

FIG. 14B is a second cross-sectional illustration of the fungal materialbeing pierced with a pointed element descending vertically therethrough;

FIG. 14C is a third cross-sectional illustration of the fungal materialbeing pierced with a pointed element descending vertically therethrough;

FIG. 14D is a fourth cross-sectional illustration of the fungal materialafter being pierced with a pointed element descending verticallytherethrough;

FIG. 15A is a first cross-sectional view of fungal material and voidspaces;

FIG. 15B is a second cross-sectional view of fungal material and voidspaces;

FIG. 15C is a third cross-sectional view of fungal material and voidspaces;

FIG. 15D is a fourth cross-sectional view of fungal material and voidspaces;

FIG. 15E is a fifth cross-sectional view of fungal material and voidspaces;

FIG. 15F is a sixth cross-sectional view of fungal material and voidspaces;

FIG. 15G is a perspective view of fungal material and void spaces;

FIG. 16A illustrates part one of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16B illustrates part two of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16C illustrates part three of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16D illustrates part four of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16E illustrates part five of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16F illustrates part six of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16G illustrates part seven of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16H illustrates part eight of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16I illustrates part nine of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16J illustrates part ten of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 16K illustrates part eleven of an eleven-part alternative methodinvolving agitation of the fungal material;

FIG. 17A illustrates a first of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17B illustrates a second of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17C illustrates a third of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17D illustrates a fourth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17E illustrates a fifth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17F illustrates a sixth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17G illustrates a seventh of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17H illustrates an eighth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17I illustrates a ninth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 17J illustrates a tenth of various steps involving the alterationof the fungal material via displacement of fungal material and regrowthof surrounding fungal material;

FIG. 18A is a perspective view of a first step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete fibrous elements;

FIG. 18B is a perspective view of a second step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete fibrous elements;

FIG. 18C is a perspective view of a third step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete fibrous elements;

FIG. 18D is a perspective view of a fourth step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete fibrous elements;

FIG. 19A is a perspective view of a first step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete particles;

FIG. 19B is a perspective view of a second step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete particles;

FIG. 19C is a perspective view of a third step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete particles;

FIG. 19D is a perspective view of a fourth step of an optional methodwherein the surface plane of the growing fungal material is embeddedwith discrete particles;

FIG. 20A illustrates a first step in the process of forming and fusing athree-dimensional fungal object in the form of a tube shaped around asolid cylinder, in this instance while remaining attached to a nutritivevehicle;

FIG. 20B illustrates a second step in the process of forming and fusinga three-dimensional fungal object in the form of a tube shaped around asolid cylinder, in this instance while remaining attached to a nutritivevehicle;

FIG. 20C illustrates a third step in the process of forming and fusing athree-dimensional fungal object in the form of a tube shaped around asolid cylinder, in this instance while remaining attached to a nutritivevehicle;

FIG. 20D illustrates a sectional view of the same object taking form asillustrated in FIG. 20A-20C, with nutritive vehicle, intermediate layer,cylindrical form, and overlapping and joined elements of the fungalmaterial resting upon the cylinder;

FIG. 21A illustrates a first step of a method for growing fungalmaterial in a targeted three-dimensional form (in this case ahemispherical shape), with a view of the object growing in a controlledand administrable environment;

FIG. 21B illustrates a second step of a method for growing fungalmaterial in a targeted three-dimensional form (in this case ahemispherical shape);

FIG. 21C illustrates a cross sectional view of the first step of amethod for growing fungal material in a targeted three-dimensional form(in this case a hemispherical shape);

FIG. 21D illustrates a cross sectional view of a second step of themethod for growing fungal material in a targeted three-dimensional form(in this case a hemispherical shape) illustrated in FIG. 21A-21B;

FIG. 21D illustrates a cross-sectional view of a second step of themethod for growing fungal material in a targeted three-dimensional form(in this case a hemispherical shape) illustrated in FIG. 21A-21B; and

FIG. 22A illustrates a first of multiple steps in the formation of afungal object wherein a composite material is placed on a fungalmaterial and a portion of said fungal material is delaminated from theintermediate layer, folded to encase the composite material, and fusedwith the portion of the fungal material left in contact with theintermediate layer and underlying nutritive vehicle;

FIG. 22B illustrates a second of the multiple steps detailed withrespect to FIG. 22A;

FIG. 22C illustrates a third of the multiple steps detailed with respectto FIG. 22A;

FIG. 22D illustrates a fourth of the multiple steps detailed withrespect to FIG. 22A; and

FIG. 22E illustrates a fifth of the multiple steps detailed with respectto FIG. 22A.

DETAILED DESCRIPTION OF THE INVENTION

The forthcoming descriptions of the present invention have beenpresented for the purpose of illustration and description. It is notintended to be exhaustive or to limit the invention to the precise formdisclosed. Many modifications and variations are possible in light ofthe above teachings. It is to be understood that other embodiments maybe utilized and changes may be made without departing from the scope ofthe present invention, and that the scope of the present invention notbe limited by this detailed description, but by the claims and theequivalents to the claims appended hereto.

Various inventive features are described below that can each be usedindependently of one another or in combination with other features.However, any single inventive feature may not address any of theproblems discussed above or only address one of the problems discussedabove. Further, one or more of the problems discussed above may not befully addressed by any of the features described below.

As used herein, the singular forms “a”, “an” and “the” include pluralreferents unless the context clearly dictates otherwise. “And” as usedherein is interchangeably used with “or” unless expressly statedotherwise. As used herein, the term “about” means +/−5% of the recitedparameter. All embodiments of any aspect of the invention can be used incombination, unless the context clearly dictates otherwise.

Unless the context clearly requires otherwise, throughout thedescription and the claims, the words ‘comprise’, ‘comprising’, and thelike are to be construed in an inclusive sense as opposed to anexclusive or exhaustive sense; that is to say, in the sense of“including, but not limited to”. Words using the singular or pluralnumber also include the plural and singular number, respectively.Additionally, the words “herein,” “wherein”, “whereas”, “above,” and“below” and words of similar import, when used in this application,shall refer to this application as a whole and not to any particularportions of the application.

The description of embodiments of the disclosure is not intended to beexhaustive or to limit the disclosure to the precise form disclosed.While the specific embodiments of, and examples for, the disclosure aredescribed herein for illustrative purposes, various equivalentmodifications are possible within the scope of the disclosure, as thoseskilled in the relevant art will recognize.

For the specification, the following terms shall apply.

Administrable space: A volume in which the orientation of fungal hyphaecan be directed, e.g. through physical manipulation, chemicalmanipulation, or phototropic, skototropic, or optogenetic control.

Composite material: Any material not the same as a fungal tissue thatcomes into contact with said fungal tissue. Composite material may thusconsist of fungal tissue.

Growth pattern of hyphae: The micro- and macro-structures of thematerials that make up the hyphae, e.g. the arrangement of chitinmolecules, the thickness of hyphal walls, the number of branches, theamount of hyphal fusion, and the orientation of the hyphae

Hyphae: The branching filaments that make up a fungal tissue.

Hyphal fusion: The biological process by which fungal hyphae join, alsoknown as hyphal anastomosis. A form of fused growth.

Nutritive vehicle: A substance capable of providing nutritionalresources for the growth and metabolic processes of a fungal tissue.

Porous material: A material with any number, shape, and orientation ofzones permeable to fungal growth but not to the nutritive vehicle.

Turning first to FIG. 1, a block diagram of a method of producing afungal structure for industrial applications in accordance with thepreferred embodiment of the present invention is illustrated. One methodof making the fungal material structure 80 includes providing anutritive vehicle 30 having a mixture of discrete particles andnutrients 20 in an enclosure. A fungal inoculum 10 is prepared, which ismade up of a desired fungi strain, which can be any vegetative, sexual,or asexual structure of a fungus that is capable of growing a new fungalcolony. The fungal inoculum 10 needs the nutrients 20 to grow, and whichare blended with the nutritive vehicle 30. Thus, the fungal inoculum 10is introduced into the nutritive vehicle 30 within the enclosure, whichprovides an even distribution of the fungus throughout. An intermediatelayer 40 is placed on a surface of the nutritive vehicle 30, providing amedium through which fungal hyphae might grow or be prohibited thereof50 and preventing the expressed fungal material 50 from permanentlyadhering to the nutritive vehicle 30. Live fungal material 50 is grownfrom the fungal inoculum 10 on the nutritive vehicle and through theintermediate layer, and grows in a direction away from the exposed faceof said nutritive vehicle. 30. An arrangement of fibers is optionallyplaced on the surface of the newly grown fungal material atop theintermediate layer as shown at block 45. Live fungal material isdirected to grow through these fibers and form a composite material withenhanced material properties. Throughout growth, live fungal material 50is manipulated as indicated at block 60 to achieve a desired thickness,shape, size and quality of fungal material. The intermediate layer 40 isdelaminated as indicated at block 70 to terminate further growth of thefungal inoculum 10, thereby obtaining the fungal material 80, which canbe processed for further use as indicated at block 90.

In FIG. 2, a flowchart 100 illustrates the method of producing thefungal material for industrial applications in accordance with thepreferred embodiment of the present invention. The method comprises thesteps of: obtaining a fungal inoculum comprising a desired fungi strainas indicated at block 102, placing a nutritive vehicle within anenclosure as indicated at block 104, inoculating the nutritive vehiclewithin the enclosure with the fungal inoculum as indicated at block 106,placing an intermediate layer on the top surface of the nutritivevehicle within the enclosure as indicated at block 108, placing anoptional additional material on top of the intermediate layer and/ormycelial layer, throughout which fungal material can grow to form acomposite material, stimulating the growth of the fungal inoculum into adense network of hyphae and allowing the hyphae to produce the fungalmaterial on the nutritive vehicle and the intermediate layer asindicated at block 110, periodically manipulating the growth of thefungal material to grow the fungal material with desired characteristicsas indicated at block 112, delaminating the intermediate layer from thenutritive vehicle to obtain a fungal material structure as indicated atblock 114, and processing the fungal materials for various industrialapplications as indicated at block 116.

Nutritive Vehicle

The nutritive vehicle 30 includes discrete particles and nutrients thatallow the desired fungi strain to grow over a period of time bydigesting the nutrients. The nutritive vehicle 30 is any materialadequate to provide for the growth of the fungal material 50. Anutritive vehicle 202 can be a ligno-cellulosic material withappropriate pH balance and other nutrients commensurate for thepropagation of a desired fungi strain.

Fungal Inoculum

A fungal inoculum is an amplifiable colony of a desired fungal strain.The fungal species is preferably selected from the fungal kingdom orderPolyporales, the Family Ganodermataceae, with preference to theGanoderma lucidum, Ganoderma tsugae, Ganoderma applanatum, Ganodermaresinaceum, Ganoderma oregonense. Other preferred candidate includeTrametes versicolor, Trametes pubescens, Schizophyllum commune, andPolyporous squamosus. The desired fungi strain is propagated throughoutthe nutritive vehicle so that it is fully colonized by the fungalmycelium. In one method, the fungi may be propagated by application topasteurized nutritive vehicle within sealed plastic bags with microporemembranes (for atmospheric regulation).

Growth Enclosure

A growth enclosure can be any type of container adapted for culturingcells, bacteria, and/or mycelia and that can contain the nutritivevehicle while minimizing opportunities for infection and allowing forthe control of environmental factors such as temperature, humidity,light levels, and CO2 and O2 concentrations.

Fundamental is that the enclosure controls air exchange and O2,directing the growth to only specific areas through access toatmosphere, e.g. through the intermediate layer.

The enclosure is used to contain the nutritive vehicle. In oneembodiment, the nutritive vehicle 30 is uniformly distributed in anenclosure and leveled to substantially form a plane normal to thedirection of the fungal materials growth 50, aiding in the even anduniform growth of resultant planar fungal material.

The inoculum of the desired fungi strain 10 can be partially or fullycolonized throughout the nutritive vehicle 30 within the enclosure.

The enclosure and the exposed surface of the nutritive vehicle 30 may beformed into one of or any combination of planar and non-planar surfacesto determine the shape of the resultant fungal material.

Intermediate Layer

The intermediate layer 40 is placed on an exposed surface of thenutritive vehicle 30 and is intended to physically isolate the growingfungal material from the nutritive vehicle. It can take the form of amembrane or fabric that is permeable to the growing fungal material 50but not to the particles of the nutritive vehicle. It enables uniformgrowth of the fungal material 50 by providing uniform initial conditionsof growth and enables the fungal material 50 to be cleanly removedwithout damaging the nutritive vehicle 30 during the delaminationprocess 70.

The intermediate layer 40 facilitates uniform separation of the fungalmaterial 50 from the nutritive vehicle 30 by controlling the interactionof the nutritive vehicle with the fungal material. Thus, theintermediate layer 40 prevents the fungal material 50 from permanentlyadhering to the nutritive vehicle 30, and damaging or tearing of thenutritive vehicle 30 when removing the fungal material 50. The nutritivevehicle 30 can thus be reused to grow additional mycelial structures.Said again, when the living fungal tissue has been removed from thenutritive vehicle from which it has grown, it can be reattached to thesame or similar nutritive vehicle, then allowed to reform as a result ofnatural bonding and fusing of fungal hyphae. This can be a somaticclonal type, or might be differentiated sexually, or by differentspecies. The intermediate layer 40 may be fully or partially permeableacross its surface. Growth of fungal material will be blocked inimpermeable areas, allowing for masked or patterned growth.

In one embodiment, the intermediate layer is a stand-alone componentseparate from the enclosure and the nutritive vehicle. In anotherembodiment, the intermediate layer is attached to or is permanently apart of the enclosure. In another embodiment, the intermediate layer 40is embedded within the nutritive vehicle 30.

The material of the intermediate layer 40 may comprise lignin or otherbiodegradable compounds to interact with the mycelium 50. In analternate embodiment, the material of the intermediate layer 40comprises a polymer that is not degraded by the fungi.

In certain embodiments, the intermediate layer is formed in situ. Inthese embodiments, an intermediate layer can be created on top of thecolonized nutritive vehicle through heat or chemical polymerization ofthe cellulose, chitin and other components of the nutritive vehicle. Inan alternative embodiment, a thin initial layer of mycelial growth onthe exposed surface of the nutritive vehicle can be modified by physicaltending of the growing tissue or by applications of heat, chemicaltreatments, and/or powders, gels, or other materials to create anintermediate layer.

The intermediate layer may be designed to accept controls by variousmeans (including electrical actuation) to allow for the application ofdynamic filtering functions to growing fungal tissue

Controlling Growth

The fungal material 50 may be periodically manipulated 60 to directgrowth in ways that confer desired characteristics, including density,evenness, and higher strength. Without careful tending, the fungalmaterial 50 will differentiate into sclerotic tissues, primordium,fruiting bodies, and other tissues. It is critical to maintain controlthrough careful manipulation to obtain uniformly grown mycelialstructures with desired characteristics. The periodic alteration ofmorphology in the apical extension of a colony of growing fungal hyphaeacts as an inhibiting agent on localized developmental pathways of thattissue.

The growth of the mycelium 50 is directed by direct means and passivemeans. The direct means of manipulations 60 comprise intentional changesthrough the application of pressure, ablation, abrasion, cutting,chemical additives, and/or electromagnetic stimulation and othermethods. The passive means of manipulations 60 may be achieved throughcontrol of environmental conditions and/or the composition of thenutritive vehicle. In the preferred embodiment, both means ofmanipulation 60 are used.

Various growth compounds and fungal hormones exist, such as10-oxo-trans-8-decenoic acid (ODA), that can be used to change growthcharacteristics as is desired. Chemical supplements, such as forskolin,may also be added to the nutritive vehicle, or misted onto the surfaceof growing fungus materials. Finally, growing the material through anintermediate layer with a controlled electric field can allow one toinduce, prohibit, or otherwise act to determine characteristics ofgrowing tissues. All of the techniques can be used to address and alterthe growing material and its final characteristics.

The orientation of fungal hyphae changes the morphology of the tissueand thus the mechanical characteristics of the material. As the tissuegrows in periodic layers, it is possible to orient the growth of layersin succession to thus create structures at microscopic and macroscopiclevels of the fungus material composite, and to produce differentmaterial characteristics as a result.

The composition and growth habits of growing hyphae can be directed byvarious means. For example, protein inhibition and/or osmotic shock canincrease the normal synthesis of chitin, which could have significantimplications for material strength.

The environment can be controlled in many ways to affect local andglobalized growth of the fungal material 50, including through thestructure of the enclosure and any lid applied to the exposed surface.In one instance, the control of light frequency and intensity isessential as a means of control.

In the instance that intentional inconsistencies are desired within theproduct, disturbances across the plane of growth can be induced throughenvironmental controls and application of various physical and chemicaltreatments. In one embodiment, differing environmental controls areapplied to particular regions of the growing material to create specificdesired and localized effects. For example, the relative concentrationsof gaseous O2 and CO2 can be used to create desired growth habits. Inanother instance, control of temperature can be used to similar effect.In another instance, aspirated air applied to areas of the growingsurface can be used to prevent or promote certain developments of thegrowing fungal organism.

Aspects of the colonized nutritive vehicle might be intentionallyorganized to control the growth of the fungal tissues, including thedistribution of nutrients within the nutritive vehicle, the intentionaluse of antibiotics and other agents for masking, and the application ofother amendments to inhibit and/or promote various types of growth.Other environmental responses of the organism (gravitropism,phototropism, etc.) may be used to control growth, both through activecontrol of the growth environment, or by passive control from the designand organization of the growth environment.

The manipulation 60 of the fungal material 50 can be performed byphysical means. In one instance, the hyphae are flattened and laid inone direction using a roller. The roller depresses the hyphae into aplanar form. When the hyphae regrow, they express an arbuscular form,and the rolling method is used to weave the body of the fungal material50 into novel patterns. Due to such physical manipulation 60 of thefungal material 50, the hyphae can be grown into particular anddetermined directions such that they can be arranged into orthogonalstructures, lattices, and other two-dimensional and three-dimensionalorganizations. With the consistent and patterned manipulations 60 ofhyphal growth with this method, the fungal material 50 can be formed inlayered structures with determined arrangements of fungal tissue (e.g.alternating layers with orthogonally-arranged fibers). In addition todetermining the structure of the hyphal network, this form ofmanipulation also homogenizes the mycelial tissue by deterring it fromdifferentiating and developing primordia or other tissues.

In another embodiment of physical manipulation, the mycelial tissues maybe periodically scratched and/or furrowed to create a fungal materialstructure with highly intergrown layers. The resulting entanglementbetween adjacent layers and the high surface area of the interlayersurface contributes to resistance to delamination between individuallayers.

In another embodiment, the growth of the hyphae is stimulated by lightat the macroscopic level. In yet another embodiment, the growth of thehyphae at the cellular level is controlled by the use of optical signalsand fungal strains that are genetically-modified to respond to suchsignals.

In an alternate embodiment, water, liquid solutions, gels and otherviscous agents may be deposited on the growing surface of the fungaltissue in determined volumes and distributions. These agents affectlocalized conditions by encouraging or inhibiting the growth andextension of fungal hyphae. Through this deposition of viscous agents,localized and emergent tissue development can be affected withdetermination and fidelity. Hydrostatic charges on said depositedviscous agents also have influence over the behavior and direction ofapically extending hyphae above the tissue surface.

Growth of Composite Materials

In an alternative embodiment, to direct growth and/or produce compositematerials, materials may be incorporated into the growing fungal tissuewhile the fungal material is still viable. In one embodiment,cellulose-based, synthetic or other organic fibers including varioustextile forms (e.g. woven, knit, fulled, felted) of preferred lengthsand structural characteristics are deposited on the exposed surface ofthe growing fungal tissue, allowing for the growth of a compositematerial. The composition and organization of the composite fibersenables the fungal tissue to be engineered, enhancing mechanicalproperties of the overall material including tensile and compressivestrength.

In an alternate embodiment, fungal tissue can be grown through 2D and 3Dmatrices and objects of various materials to create composites withdesired characteristics and qualities. This added layer of material maybe composed of any material that fungal cells can grow through (poresize larger than 1 micron). These layers may be pressed onto or near thesurface of the growing cells or otherwise impressed upon its surface, orplaced between two or more layers of growing fungal material, such thatthese reinforcement layers are then incorporated into the fungal tissue.

A further laminate may be bound to the fungal tissue that has grown intoreinforcing elements and layers by attaching said laminate to thegrowing composite, which enables a method of binding all the layerstogether as a result.

Delamination

During the delamination process 70, it is possible to pull the fungalmaterial 50 off the intermediate layer 40 in a manner that theintermediate layer 40 remains adhered to the nutritive vehicle 30 andprovides subsequent secondary and further growth of the fungal material50. Thus, multiple sheets of fungal material 50 can be produced duringthe process.

An alternative embodiment of the invention involves the reapplication ofthe delaminated fungal material 70 onto the same nutritive vehicle 30,but in a different geometric orientation. Yet another instance involvesthe reapplication of the delaminated fungal material 70 onto a nutritivevehicle inoculated with a different type of fungus. The produced fungalmaterial 80 can then be processed for use by various methods describedbelow.

Fused Growth

Hyphal fusion is one mechanism by which fused growth can occur. It isthe biological process by which distinct and compatible fungal hyphaejoin, also known as hyphal anastomosis. This process can be stimulatedand directed for the production of novel materials.

Living mycelial sheets may be used as-is, or the sheet and any number ofsheets can be joined with portions of itself or other living fungalmaterial structures by stimulating growth with the applications ofpressure and availability of nutrients.

In a preferred embodiment, multiple living sheets are stacked togetherand set with a weight or pressure such that at least one sheet orportion thereof is in contact with a mass of nutrient nutritive vehicleand is incubated until these components fuse into a cohesive structure.

In an alternate embodiment, living mycelial structures may be shaped orformed into 3D objects and incubated until these components compose auniform object or fuse together in a desired way.

In one instance, any living fungal material 50 can be delaminated fromits nutritive vehicle and combined together with another structure ormany other structures, such that in some cases the fungal tissueself-adheres, which causes fusing at a cellular level. In one instance,many structures may be combined to create complex self-adhered shapes orsheets. Alternatively, areas of a single sheet of fungal material 50 canbe joined together to generate a 2D or 3D form.

For fusing to occur, the living tissues must have access to nutrients tofuel growth. In one embodiment, this can be achieved by leaving thematerial partially attached to the nutritive vehicle from which it hasbeen generated, arranging it so that the areas desired to fuse togetherare made to come into contact with one another, applying pressuresufficient to produce close contact between the areas, and allowing thematerial enough time to grow a complete bond.

In one instance of the invention, a sheet of fungal tissue is grown bymeans of fungal hyphae expressed through an intermediate and perforatedlayer into the form of a layer within a growing environment. Thisenvironment and hyphae are administrated to create a fungal material orcomposite of a desired thickness and order, upon which a part or half ofsaid sheet of material is delaminated from the intermediate layer, andfolded in such a manner that the delaminated aspect of the layer is putinto as close contact as is possible with the fungus tissue stillconnected to the intermediate layer. If such an altered version of thegrowing artifact is left for a period as short as 24 hours, the hyphaefrom the top part of the fungal tissue material will fuse to the hyphaeof the layer beneath it, growing into a joined mass.

Additionally, scoring, pricking, manipulating the surface or otherwisealtering the surface qualities of the areas to be fused may alter themechanical properties of the produced material, such as by increasingthe bonded strength of multiple laminated sheets of material, creatingvoid spaces, lattices, and other internal artifacts structured throughsuch intentional growth.

In another embodiment, non-fungus materials can be fused between fungusbased materials to serve the purpose of structural reinforcements orother modifications to the performance and aesthetics of the fungusbased materials.

Post-Growth Processing

The sheet of fungal material may be treated in such a manner as to makeit pliant and resilient towards the uses of industry. Without suchprocessing, the resultant material can be highly brittle, weak, andgenerally of low utility. The processing can be performed by drying,providing mechanical stress, and adding plasticizers, water-proofingagents, cross-linking agents and other additives. The fungal materialmade in accordance with the invention may be subjected to furtherprocessing steps to achieve a desired final product for variousindustrial processes and applications.

If left untreated, the material can become brittle when dry. Should aflexible dry state be desired, the material can be treated with abiodegradable plasticizing agent and/or worked mechanically throughoutthe drying process. In one embodiment, the plasticizing agent can be oneof any number of polyols typically used as plasticizers for chitin- andchitosan-bearing materials, such as propylene glycol. Such plasticizerscan be applied to the material by rubbing, spraying, or soaking with orwithout dilution by water or other solvents. The material may also berendered flexible by working it mechanically throughout the dryingprocess with or without the application of oils, plasticizing agents, orother additives.

The material may also be chemically cross-linked at the molecular levelto improve material qualities such as tensile strength and flexingendurance. In one embodiment, this is achieved through the applicationof tanning compounds (such as pyrogallols and catechols) and aldehydecompounds (such as glutaraldehyde). In another embodiment, this isachieved through the cross-linking between amine groups in the fungalchitin and/or chitosan molecules.

The material can be pressed with or without heat to obtain desiredthicknesses and surface qualities. In one embodiment, the material isdried and pressed between heated plates at temperatures greater than 200F to compress the material, achieve an even surface, and impart adesired surface texture.

The material can be treated with waterproofing or water-resistanceagents before, during, or after pressing, or at any stage in theproduction of the material to impart greater water-resistance to thematerial and greater water-fastness to the properties of the material.In one embodiment, the material is coated in a paste made from a mixtureof natural oils and waxes and heated with or without pressure to achievepenetration.

At various points in this process, the material may be dyed or otherwisefinished aesthetically as desired. Dyes, for example, can be appliedbefore, after, or during the plasticizing step through painting,spraying, or soaking with or without the presence of water or othersolvents. In another embodiment, the surface textures can be altered byabrasion, rolling, or other means.

The following examples reflect certain embodiments of the invention. Themethod components in the examples may be presented as steps; however,unless specifically stated, any one step need not occur before or afterany other step. Furthermore, in certain embodiments of the invention,certain steps may be omitted or repeated.

Example 1—The following example describes one method for producing aflat sheet of fungal material with an embedding composite material forreinforcement.

Step 1—Obtaining inoculated nutritive vehicle—The nutritive vehicle mayconsist of at least one of straw, hay, hemp, wool, cotton, rice hulls,recycled hardwood/softwood sawdust, water, calcium carbonate, nitrogen,sugar rich grains, and shrimp shells.

A given nutritive vehicle of a combination including the above materialsis pasteurized and let to cool to room temperature. A tissue culture ofthe desired fungi strain is introduced and left to propagate throughoutthe nutritive vehicle. The pasteurization process eliminates or limitscompetition as the fungus is colonizing the nutritive vehicle. This stepis referred to as the colonization of the nutritive vehicle.

Step 2—Loading nutritive vehicle into growth enclosure—Showing apreferred embodiment, FIG. 3 illustrates a box-like enclosure 200 forplacing a nutritive vehicle in accordance with the preferred embodimentof the present invention. The box-like enclosure 200 may in oneembodiment comprise a plywood box measuring 3′×3′×8″, which is linedwith 3 mil plastic sheeting. In one embodiment of the present invention,the enclosure 200 consists of a single compartment. In anotherembodiment of the present invention, the enclosure 200 comprises aplurality of compartments.

FIG. 4 illustrates the nutritive vehicle 202 placed within the enclosure200 in accordance with an embodiment of the present invention.

In one embodiment, 60 liters of colonized nutritive vehicle is placedinto the enclosure. The nutritive vehicle is evenly distributed at adepth of 4-6″, and flattened with pressure such that the top surface ofthe nutritive vehicle forms a flat and even surface.

Step 3—Application of intermediate layer—A top surface of the flattenednutritive vehicle 202 is covered by an intermediate layer 204 which is apermeable membrane. FIG. 5 illustrates the intermediate layer 204 placedon the top surface of the nutritive vehicle 202 in accordance with thepreferred embodiment of the present invention.

In a preferred embodiment, a prepared intermediate layer is placed inthe bottom of the empty enclosure, inoculated nutritive vehicle ispacked on top of this layer and is flattened with pressure, filling upthe enclosure to the proper depth and density. The enclosure is coveredwith a lid while the nutritive vehicle is given 2-4 days to growtogether into a mass. Finally, the solidified fungal material is takenout of the enclosure, flipped over to reveal the intermediate layer nowon the top surface, and re-placed into the enclosure, returning the lidand is left to grow.

In an alternate embodiment, the intermediate layer 204 is stretched on aframe to create a flat and tensioned surface. This tensioned fabric isplaced against a rigid surface, and the nutritive vehicle is then packedagainst the fabric to create a flat nutritive vehicle surface closelymated to the intermediate layer. In this instance, the exposed framedintermediate layer may be revealed on the bottom of the enclosure. Inanother instance, the exposed intermediate layer may be revealed on thetop of the enclosure.

Step 4—Applying composite material—In a preferred instance, theenclosure is placed in an environment conducive to growth, and fungaltissue grows through the intermediate layer after a few days. Thisgrowth is visible as a ‘fuzzy’ layer beginning to appear on the surfaceof the exposed intermediate layer. Once a visible layer of ‘fuzzy’fungal material has uniformly established itself through and on top ofthe intermediate layer (˜1-4 days), fibers and/or textile that have beenpasteurized are placed on top of this growth above the intermediatelayer. This is called a ‘composite layer’. Cellulose-based materials[e.g. cotton or rayon] are often used as they are both biodegradable anda non-preferred food source for Ganoderma, meaning they maintainstrength through the growth process in ways that lignin-bearingmaterials do not. Lignin is a preferred nutrient to this organism overcellulose. As the fungal material grows under specific environmental andphysical controls, the filamentous fibers will grow through thiscomposite, away from the colonized nutritive vehicle from which theyhave grown, and expanding further to grow another layer of hyphae. Thisbehavior of strands of hyphae growing away from a nutritional source inan expansive and explorative manner is typical to many types offilamentous fungi, and is almost universally observed in saprophyticfungi.

Various forms of textiles may be used as composite materials. Inparticular, woven or knit cellulose-based materials (e.g. cotton) withheavily fulled surfaces are preferred. The woven or knit structuresprovide strength and the tousled fibers of the fulled surfaces anchorthe fungal material to the textile, lowering the risk of delamination inthe finished composite materials.

In an alternate method for applying the intermediate layer, a sheet oftightly woven fabric is pasteurized and layered on the top surface ofthe flattened nutritive vehicle 202, such that it is pressed or sealedusing liquid to fabric and nutritive vehicle are brought as into contactas closely as possible. This is pressed so to further enhance flatnessand evenness of the top plane of interaction on the flattened layer 202.The sides of the fabric are tucked along the edges of the box-likeenclosure 200 to further assist in flattening the fabric.

In an alternate embodiment, any number of composites may be applied inany order or sequence, or between layers of fungal material, to createsophisticated and specific material qualities.

The growing fungal tissue extends a dense network of hyphae 210 throughopenings of the intermediate layer 204 of the woven fabric up into thearea above the intermediate layer 204 in an undifferentiated arbuscularmass, giving the appearance of fuzzy/fluffy white fibers. FIG. 8illustrates the first dense network of hyphae 210 growing off of thenutritive vehicle 202 through the intermediate layer 204 in accordancewith the preferred embodiment of the present invention.

After four days, the dense network of hyphae 210 forms into a livefungal tissue mat that has grown as an extrusion through theintermediate layer 204.

Step 5—Controlling environmental conditions—Growth environment ismaintained such that mycelia tissue creation is maximized. A lid 206 isplaced on top of the box-like enclosure 200 that is large enough tocontain it from the larger environment within which it is embedded. Thislid 206 preferably comprises a plurality of openings 208 and permeableaspects that allow for the transpiration of gases and humidity betweenthe inside of the growing box-like enclosure 200 and the outside. FIG. 6illustrates the enclosure lid 206 with the plurality of openings 208 inaccordance with the preferred embodiment of the present invention.

In one embodiment, a piece of acrylic plastic or other non-reactivematerial measuring 38″×38″, with the plurality of openings drilled everyfour inches in a grid array, is placed on the top of the box-likeenclosure 200 and the growing nutritive vehicle 202.

The growing box-like enclosure 200 of the covered nutritive vehicle 202is closed with the lid 206 and kept in an environment that hascontrollable variables. FIG. 7 illustrates the enclosure 200 closed withthe lid 206 in accordance with the preferred embodiment of the presentinvention. Proper atmospheric conditions inside the box include humidityranging between 20-100% RH, rich in oxygen and a temperature between50-95° F.

The room that the box-like enclosure 200 is growing within is preferablykept under positive pressure, ambient humidity levels at 10-15%, atemperature range of between 22-25° C., and total darkness. Thematerials are left to grow for a time period lasting at least two weekswith daily tending of the growing material, including manipulation ofthe growing fungal material.

Step 6—Manipulation of the growing fungal material—The growing hyphaeare manipulated in such a way to depress them on top of the intermediatelayer 204 through which they are growing, such that they are altered tobe in contact with the surface 204.

In a preferred instance a small rolling pin may be used to roll over atop surface of the intermediate layer 204 in a defined direction, thusaltering the geometry of the growing plan of extending the fungalmaterial. The entire surface of growing hyphae above the intermediatelayer 204 is treated as such to achieve a uniform surface of flattenedhyphae. FIG. 9 illustrates the first dense network of hyphae 210 shownin FIG. 8 being rolled flat in a first direction on the top surface ofthe intermediate layer 204 to form a first flattened fungal materialstructure 212 in accordance with the preferred embodiment of the presentinvention. FIG. 10 illustrates a second dense network of hyphae 214regrown from the intermediate layer 204 having the first flattenedfungal material structure 212 shown in FIG. 9 in accordance with thepreferred embodiment of the present invention. FIG. 11 illustrates thesecond dense network of regrown hyphae 214 being rolled again in asecond direction on the top surface of the intermediate layer 204 toform a second flattened fungal material structure 216 in accordance withthe preferred embodiment of the present invention. FIG. 12 illustrates athird dense network of hyphae 218 regrown from the intermediate layer204 having the second flattened hyphae 216 shown in FIG. 11 inaccordance with the preferred embodiment of the present invention.

A formula comprising Tea Tree oil, soap and water is sprayed on top;however, combinations of those liquids or just one of those liquids maybe used. This action is preferably repeated each day for the next tendays, with the direction of the roller alternating 90 degrees each day.By this method, conjoined layers of orthogonally-oriented hyphae arecreated.

In an alternate embodiment, multiple number of fungal material sheetsand a multiple number of textiles may be stacked and incubated untilthese components compose a uniform solid. In between these layers,‘scoring’ or ‘scratching’ the surfaces much like in traditional ceramicspractices may increase the strength of the bond and improve theresistance to delamination between layers.

Environmental factors within the growing box-like enclosure 200 or theroom that contain the box-like enclosure 200, may be altered so as toaffect the growth characteristics of the fungi strain. Sprays,particles, fibers, and/or other additives and solutions may be added tothe growing fungal mass. The layers might be scratched and otherwisealtered to interweave/intermesh multiple growth layers or void spaces.

Step 7—Delamination of sheet—When the extruded fungal material structurehas accreted to a desirable consistency and thickness, a sheet of fungalmaterial structure is delaminated from the intermediate layer 204 anddisassociated from the nutritive vehicle 202 from whence it originated.

A surgical knife or scissor may be used to cut through the growingsurface, close to the sides of the box-like enclosure 200 in which thegrowing mass is contained. The sheet of fungal material structure ispulled up from a corner, much as one pulls a sticker off of its paperbacking. In such a manner, the sheet of fungal tissue can be pulled offand retain its structure as an autonomous object in the form of a sheet.

In an alternate embodiment, the edge of the fungal material sheetextends over the enclosure such that no cutting is needed to separatethe sheet of fungal material from the delamination layer.

Step 8—Post-growth processing of sheet—The material is soaked in asolution of catechol tannins equal in concentration to those used inanimal leather processing. This strengthens the material throughchemical tanning. In one embodiment, this is completed by soaking thematerial in a tannin solution of quebracho tree extract at boilingtemperature for at least 4 hours.

The delaminated sheet can be treated with a plasticizing agent to impartflexibility when dry. It is soaked in a bath of 4:1 water-to-propyleneglycol for 60 minutes, optionally pressed to remove excess liquid, andthen hung to dry.

Once dry, the material is pressed at a temperature above 200 F betweensmooth heated plates at a pressure above 5 pounds per square foot for atime greater than 15 seconds. This compresses the material, imparts aneven thickness, and gives a smooth surface texture. In an alternateembodiment, the plates used for pressing the material can be texturedand used to impart a desired texture to the material.

The material can be dyed (for example by painting with an oil-basedleather dye) and dried. It can then be coated in a finishing wax(composed for example of coconut oil, carnauba wax, and beeswax) andbuffed.

The material can be treated with a water-resistance or waterproofingagent. For example, the material can be sprayed with any number ofdurable water repellant finishes such as Parylene C.

Example 2—A process for making a fungal tissue composite in accordancewith certain embodiments of the invention includes.

Step A—Nutritive Vehicle Preparation

A.1—mixing of nutrient components and water

A.2—sterilization or other reduction of biotic load

A.3—inoculation of nutritive vehicle with mushroom tissue

A.4—mixing of all components

Step B—Vessel Preparation

B.1—filling the vessel with the prepared nutritive vehicle

B.2—leveling the surface of the nutritive vehicle

B.3—preparation of intermediate layer

B.4—placing a lid on the vessel with appropriate environmental controls

Step C—incubation and growth of the fungal material

C.1—growth of fungal hyphae through intermediate layer

C.2—manipulation of hyphae as they are growing

C.3—placement of fibers, mesh, or other materials on top of growinghyphae

C.4—manipulation of hyphae as they are growing

C.5—the environmental conditions may be altered to elicit desired growthbehaviors

C.6—various additives may be brought into the surface of the fungaltissue composite or into the environmentally administrated space so asto alter the morphology and character of the growing hyphae

Step D—Delamination of the Fungus Tissue Material from the NutritiveVehicle

Step E—Manipulation and Growth

E.1—compressing the fungal tissue composite to the desired dimensionsand density

E.2—bringing one part of the fungal tissue in contact with another partof the tissue

E.3—bringing one part of the fungal tissue in contact with anothertissue element

E.4—forming the fungal tissue into a desired shape

E.5—bringing one part of the fungal tissue in contact with anotherelement

E.6—let grow for 0-96 hours

Step F—Additional Processing

F.1—drying of the fungal tissue composite, for example by convection,conduction, microwave, freeze drying or other

F.2—dried material is sanded, cut or milled to shape

Step G—Post Processing

G.1—adding a plasticizer or other post-growth processing steps

Example 3

The following Example will provide various methods and techniquesillustrated in FIG. 13-FIG. 19. This is a process through which hyphaefrom defined species of filamentous fungi are propagated from acolonizable nutritive vehicle that has been inoculated with said chosenfungi. Preferred species include the Ganodermas, the order Polyporalesgenerally, and including all saprobic types of fungal candidates thatderive sustenance from lignin and cellulose rich sources. The inventionutilizes an intermediate membrane layer, located between the colonizablenutritive vehicle and the environment into which the fungal hyphal willgrow. This intermediate layer can act as both a structural component aswell as a means of manipulating the fungal tissue that grows through it,and is instrumental in the delamination and physical removal of thedesired fungal tissue that has grown upon the colonizable nutritivevehicle.

The physical characteristics of growing fungal tissue are manipulated bydirect and passive means. Direct means include intentional and specificchanges through application of acute physical actions upon the nutritivevehicle such that its consistencies, orientations, and other physicalaspects might be altered as is desired. These include mechanical,pneumatic, hydraulic and other actions that might be brought to bearupon said fungal tissue in such a manner as to cause consequentialalteration of the fungal tissue. Other factors used to manipulate thecharacteristics of the growing fungal hyphae include: the application ofboth global and localized conditions through deposition of fluids,fibers, and other additives; control of optical conditions the growingfungal hyphae are subject to, and the selective application of heatingand cooling elements. (e.g.: a heated roller or laser can be used totransform a selected area into a hardened or otherwise artifactual stateof being). Passive means of altering the growing fungal tissue includethe control of environmental climates and conditions of the entirefungal tissue or microclimates of regions of the growing fungal tissue.

In one instance, the nutritive vehicle is placed into a frame orcontainer, the nutritive vehicle is colonized with fungal tissueexpressing fungus of a desired type, the nutritive vehicle is uniformlydistributed in a container and leveled to form a plane horizontal to theapical direction of the growth of fungal tissue above said plane. Thisplane is defined as the point of interaction or plane of interaction.Next, an intermediate solid or woven membrane, fabric or mesh is placedatop the plane of interaction so as to occupy the plane. Such a materialcan enable uniform growth.

Environmental conditions are maintained to a standard necessary forgrowth and extension of hyphae. Next, the fungal material isperiodically manipulated to grow in particular and determined directionssuch that it can be arranged into orthogonal structures, lattices, orother layered organizational arrangements. In such a way, distinctlayers can be induced to grow with determined states and structuralformations within the body of the fungal tissue.

Once the desired thickness and organization is achieved, the fungaltissue is delaminated from the intermediate layer. The presence of theintermediate layer enables the fungal material to be cleanly removedwithout affecting the colonizable nutritive vehicle from which itpermeated. The sheets or materials can be removed in such a way that theintermediate layer remains as a part of the generative base and a newskin of fungal tissue can once again grow through it in a permeablemanner. The fungal material is then processed to product specifications.

In Detail

-   1. This disclosure requires a volume of nutritive vehicle that is    colonized with a chosen of fungus of interest that has the habit or    ability to generate fungal tissue within said instance as is    desired. Not all fungus will express fungal tissue in the manner    necessary for the invention, and certain organisms function better    than others.-   2. In one instance, a container may be used to hold the inoculated    nutritive vehicle. The material does not have to be fully colonized    before entering the container in which it will be housed, but at    some point, the material may be fully colonized to properly express    tissue as is desired. In the instance that an even and uniform    product is desired, the boundary with the greater environment from    which the tissue is desired to be expressed is flattened, and the    nutritive vehicle is loaded in an even way such that there is    homogeneous distribution within its planar form. Alternative    instances utilize non-planar surfaces of nutritive vehicle,    heterogeneous distribution of nutritive vehicle or combinations of    these techniques.-   3. At a certain point in time, after the nutritive vehicle has been    inoculated, an intermediate layer, distinct from the inoculating    nutritive vehicle, is placed atop the nutritive vehicle. In one    instance, the intermediate layer is a flat surface. In one instance,    this flat surface is a piece of woven fabric laid atop the nutritive    vehicle, such that the two materials are immediately adjacent, or    such that the fabric is embedded and the nutritive vehicle and    fabric occupy the same plane. Alternatively, other membranes,    sheets, meshes, pellicles and tissues can be used as an intermediate    layer.-   4. In one embodiment, the intermediate material is a fabric of a    consistency that will not permanently bind to the fungal material,    (that is, it will only allow fungal tissue to pass through the    fabric without degradative or incorporating agencies altering it    such that it is difficult to remove the fungal tissue that has grown    upon said fabric). This fabric as an intermediate layer allows one    to separate the fungal material from the nutritive vehicle without    additional processes, and frees the nutritive vehicle to be reused    for the growth of further fungal tissues. Without the use of an    intermediate material, the fungal material will unevenly generate,    bind, and fuse to the heterogeneously ordered fibers that constitute    the nutritive vehicle, leading to a disrupted surface of the    nutritive vehicle, and unevenness in the growing sheet of fungal    tissue. In this embodiment of the invention, the presence of a    uniform and flat intermediate material atop of nutritive vehicle    enables a consistent surface for the fungal material to grow from,    and upon which to be further manipulated. Another embodiment of this    invention can be achieved without any fabric, membrane, or    substantive intermediate layer. An intermediate layer can be created    on top of the colonized nutritive vehicle through heat or chemical    polymerization of the cellulose, chitin or other materials that    constitute the nutritive vehicle or fungal tissue/    -   a. Spray on sealers, chemicals and other liquids can also be        used to create a non-woven, permeable, or solid intermediate        layer between the colonized nutritive vehicle and the fungal        tissue that will be generated from said colonized nutritive        vehicle.-   5. The remaining nutritive vehicle may then be used again to grow    new expressions of fungal tissue. Another instance involves the    reapplication of the delaminated fungal material onto the same    nutritive vehicle from which it had been removed, but in a different    orientation or formation. Another instance involves the    reapplication of the delaminated fungal material onto a nutritive    vehicle inoculated with a different type of fungus.-   6. Direct electrical stimulation and actuation of intermediate    layer.    -   a. In one embodiment of the invention, the intermediate layer        consists of system that allows for electric current to be        applied to specific X and Y coordinates, such that precise and        determined electrical current might be applied to a desired        location of said intermediate layer. Localizable electrical        stimulation is used to change composition and growth habits of        hyphae, for example increasing the frequency of hyphal fusion        and branching.    -   b. In another embodiment, an electric current in continuous or        discontinuous form is applied to charge the entire surface in a        gross manner.    -   c. In another instance, an electric current is directed toward        the actuation of characteristics of the intermediate layer, such        that a permeable membrane might alter its surface qualities to        open or close pores, or otherwise alter its surface from said        electrical stimulation to actuate a field, mechanisms, or        artifacts, or otherwise have an effect on the growth and        characteristics of the growing fungal tissue.-   7. Surface deposition, application, alteration    -   a. Different types of additives can be incorporated into the        growing fungal tissue through application or deposition. In one        embodiment, cellulose and other organic fibers of preferred        lengths, flocculation, and other structural characteristics are        deposited on the top of the surface plane of the growing fungal        tissue such that said fibers are all oriented along a preferred        axis or other orientation. These deposited fibers are        interspersed in an ordered manner as is desired across the        surface of said fungal tissue. These fibers can be used as        guides and scaffolds for the orientation of the fungal tissue,        both to promote or inhibit growth of the tissue in preferred        ways. These fibers enable the engineering of macro and micro        qualities of the fungal tissue including strength, shear,        elasticity, and memory. Alternatively, other types of fibers,        and organization of said fibers can be used.    -   b. Particles can be dispensed and otherwise broadcast and        impressed upon the surface of said growing fungal tissue.        Through deliberate and successful application of said particles,        and subsequent engulfment within said fungal tissue, the        placement of particles can be determined within a        three-dimensional matrix.    -   c. Water, liquid solutions, gels and other viscous agents can be        deposited on the growing surface of the fungal tissue in        determined volumes, patterns and distributions. These viscous        agents affect localized conditions by making available resources        that can encourage or inhibit qualities of growth and extension        of fungal hyphae. Through this deposition of viscous agents,        localized and emergent tissue development can be affected with        determination and fidelity. Hydrostatic charges on said        deposited viscous agents also have influence over the behavior        and direction of apically extending hyphae above the tissue        surface.    -   d. In another instance, a layer of nylon textile or other        materials are impressed into the surface of a fungal material,        and on top of this is placed another fungal material. Over time,        the fungal materials will fuse together, embedding the composite        material within.-   8. In the most generic term, an inoculated nutritive vehicle for a    given fungus will naturally differentiate towards the generation of    sexual organs if left unregulated. In a preferred method, the hyphae    growing above the intermediate layer are manipulated, stressed, or    otherwise altered and depressed or otherwise made to come in contact    with the surface from which they had originally extended. If treated    in such a way the fungal material will be withheld from    differentiating into organs of sexual reproduction, and instead will    return to a searching and extending manner above the surface from    which it is growing. A process of stress, alteration, and    stimulation can be repeated indefinitely to repress the fungal    organism from establishing sporulating or otherwise desirable    tissues.    -   a. Next, it is necessary to create the conditions that optimize        arbuscular expression. In one instance, a permeable intermediate        layer ensures uniformity in fungal tissue expression as it        permeates through said intermediate layer. From here on,        mycelial growth is manipulated via external manipulation and        signals, and the fungal tissue's internal expressions and        signals.    -   b. Without proper control, fungal tissue will differentiate into        tissues that will become organs of sexual reproduction and        sporulation. In the instance that an undifferentiated product is        desired, environmental conditions serve as a primary agent by        which organismal development might be determined and directed.        In the instance that intentional inconsistencies are desired in        the product, disturbances across the plane of growth can be        induced through environmental controls. In one instance, the        relative presence of O2 and CO2 can be used to determine desired        growth habits. In another instance, control of temperature can        be used to similar effect. In another instance, aspirated air        that is introduced to the growing environment can be used to        prevent or promote development of the growing fungal organism.        Other internal aspects of the colonized nutritive vehicle might        be intentionally organized to direct the growth of the organism,        including distribution and availability of nutrients within the        nutritive vehicle, intentional use of antibiotics for masking,        physical amendments and additives that might also inhibit and/or        promote various forms of growth. Other tropic responses of the        organism (gravitropism, phototropism, etc.) can be used to        direct growth, both through immediate control of the growing        environment, or by design and organization of said environmental        conditions.-   9. Alteration of the surface of the growing mycelial mass is primary    to engineering fungal tissue development and constitution.

Referring next to FIG. 13A-13G, in a preferred embodiment, the hyphaemight be flattened with a roller in one direction, as the roller crossfrom point A to point B. In doing so, the roller flattens the hyphaeback onto the plane from which they grew. As the hyphae re-growvertically, a roller can now pass from point B to point A, againflattening the vertically growing hyphae. This method can be employed toweave the body of fungal material into novel patterns that aredeliberately synthetic in organization of the orientation andconnections of the hyphae that constitute its said fungal tissue.

-   -   1. Growth of arbuscular hypha above plane of fungal tissue    -   2. Fungal material flattened in a determined direction    -   3. Regrowth of arbuscular hypha above plane of fungal tissue    -   4. Fungal material flattened in a determined direction    -   5. Regrowth of arbuscular hypha above plane of fungal tissue    -   6. Fungal material flattened in a determined direction    -   7. Cross section showing alternatively oriented directions of        fungal tissue in a determined pattern.

Turning to FIG. 14A-D, in a preferred embodiment, the fungal tissuemight be pierced or otherwise displaced with a pointed elementdescending vertically through said fungal tissue. After the element hasbeen depressed into said fungal tissue and removed, a void space iscreated as a result. Subsequent regrowth of fungal material fills invoid spaces, resulting from pointed element, and creating verticalconnection plugs between vertical aspects of said tissue (saidvolumetric space can also be filled with a secondarymaterial/amendment/additive, other fungal species, other organisms)

-   -   1. Cross section of fungal tissue with pointed element that will        be depressed vertically into said fungal tissue.    -   2. Pointed element depressed within fungal tissue    -   3. Pointed element removed from fungal tissue, leaving a void        space created as a result.    -   4. Subsequent regrowth of fungal material, which fills in void        space resulting from pointed element, and creating vertical        connection plugs between vertical aspects of said tissue (said        volumetric space can also be filled with a secondary        material/amendment/additive, other fungal species, other        organisms)

Turning next to FIG. 15A-G, this method can be employed to grow thefungal material into novel patterns that are deliberately synthetic inthe organization of their tissues through their structural constitution.Vertical and horizontal aspects of said tissue can be propagated in sucha manner to intentionally connect, weave, and otherwise engineersuccessive layers of chronologically emergent mycelial growth.

-   -   1. Cross section of fungal tissue, with pointed elements that        will be depressed vertically into said fungal tissue.    -   2. Pointed elements depressed within fungal tissue    -   3. Pointed elements removed from fungal tissue, leaving void        space created as a result.    -   4. Subsequent regrowth of fungal material, filling in void        spaces from pointed elements, and creating vertical connection        plugs between vertical aspects of said tissue (said volumetric        space can also be filled with a secondary        material/amendment/additive, other fungal species, other        organisms)    -   5. Subsequent repeat of actions in offset manner.    -   6. Orthogonal view of a section of fungal tissue treated as such        in a repeated manner, showing how patterned X Y Z configurations        of vertical and horizontal aspect so said tissue might be        determinatively organized.

Turning next to FIG. 16A-K, in a preferred embodiment, the sheet offungal tissue can be agitated by a bristled roller or other device,mechanism, or other agency that displaces and shreds the top surface ofsaid tissue as it moves across its surface from one direction toanother, to create a surface with an uneven manner. Said disturbedtissue regrows, is depressed or otherwise levelled into a form as isdesired, with repeated agitation, regrowth, and depression.Interconnecting patterns of growth between chronologically developingtissues can be determined through said methods.

-   -   1. Cross section of fungal tissue, showing distinct growth of        tissue in vertical manner.    -   2. Toothed comb or other element raked across the top surface of        fungal tissue    -   3. Shredded and otherwise agitated surface after passage of        toothed comb    -   4. Regrowth of tissue with infilling fungal material (said        volumetric space can also be filled with a secondary        material/amendment/additive, other fungal species, other        organisms)    -   5. Compression of arbuscular hypha so as to form uniform top        surface.    -   6. Regrowth of top surface into distinct layers    -   7. Repeat: Toothed comb or other element raked across the top        surface of fungal tissue.    -   8. Repeat: Shredded and otherwise agitated surface after passage        of toothed comb.    -   9. Repeat: Regrowth of tissue with infilling fungal material of        said tissue (said volumetric space can also be filled with a        secondary material/amendment/additive, other fungal species,        other organisms)    -   10. Repeat: Compression of arbuscular hypha so as to form        uniform top surface.    -   11. Cross section of fungal tissue after repeated cycles of        agitation and regrowth, showing heterogeneous intermixing and        regrowth of adjacent areas within the material.

As shown in FIG. 17A-J, the fungal tissue may be altered with an elementthat will displace said fungal material as it moves in a transversemanner across it, creating contiguous elements, pits, channels, latticesand other alterations of said tissue as a result. Said altered tissueregrows to connect or fill said displaced aspects of the transversedsurface of said fungal tissue. Interconnecting patterns of growthbetween chronologically developing tissues can be determined throughsaid methods. Woven tissues and otherwise conjoined reinforcements offungal tissue can be determined in such manner.

-   -   1. Cross section of fungal tissue, showing distinct layers, with        sharpened element that will displace said fungal material as it        moves in a transverse manner across it.    -   2. Sharpened element dislodging and altering said fungal tissue.

Two alternative options are presented

-   -   1. Fungal tissue after passage and interaction with sharpened        element, demonstrating displaced but still contiguous aspects of        said fungal tissue altered in relation to the greater fungal        mass upon which it is in contact. In another instance of this        step a roller may be moved across the top of the fungal tissue        in a transverse manner so as to impress one aspect of the        organism in greater proximal contact with other aspects of        itself. Over time the fungal distinct aspects of the greater        fungal tissue will bind and form into a contiguous whole.    -   2. Fungal tissue after passage of sharpened element from one        direction to another, demonstrating extraction of channel like        displacement of said fungal tissue.        -   i. Regrowth of tissue into channel void space with infilling            fungal material (said volumetric space can also be filled            with a secondary material/amendment/additive, other fungal            species, other organisms)        -   ii. Sharpened elements dislodging and altering said fungal            tissue from another direction.        -   iii. Fungal tissue after passage of sharpened element from            one direction to another, demonstrating extraction of            channel like displacement of said fungal tissue.        -   iv. Orthogonal view of a section of fungal tissue treated as            such in a repeated manner, showing how with patterned X Y Z            configurations of vertical and horizontal aspect said tissue            might be purposefully organized.

As shown in the preferred embodiment of FIG. 18A-D, the surface plane ofthe growing fungal tissue is embedded with discrete fibrous elementsduring its manufacture such that said fibers are all oriented along apreferred axis or other orientation. These deposited fibers can be usedas guides or scaffolds for the orientation of the fungal material, bothto promote or inhibit growth in preferred ways. These fibers enable theengineering of macro and micro qualities of the fungal tissue, includingstrength, shear, elasticity, chirality of deformation, and memory.Alternatively, other types of fibers, and organization of said fiberscan be used.

In an alternative embodiment (not shown), a discrete fibrous filament isplaced upon growing fungal tissue in a preferred geometric orientation.Then, a layer of discrete filaments is placed to form a preferredorientation. Next, additional mycelial growth encapsulates the fibrousfilaments, after which a new layer of filaments is added in a differinggeometric orientation from the previous filament layer. Finally, anotherlayer of filaments is placed upon the fungal tissue. Subsequent growthoccurs from steps 2 onward, until completion.

As shown in FIG. 19A-D, particles may be dispensed and otherwisebroadcast and impressed upon the surface of said growing fungal tissue.Through deliberate and successful application of said particles, andsubsequent engulfment within said fungal tissue, the placement ofparticles can be determined within a three-dimensional matrix.

In an alternative embodiment (not shown), a particle is dispensed andimpressed upon growing fungal tissue, a layer of discrete particles isembedded in their particular geometric orientation, and growth proceeds.Next, additional mycelial growth engulfs the discrete particles, thusincorporating them into the structure, after which a new layer ofparticles is added. Finally, the new layer is embedded in a differinggeometric orientation from the previous layer, creating athree-dimensional matrix of particles within the fungal tissue. Thesteps herein may be repeated.

Thus, although the invention has been described with respect to specificembodiments, it will be appreciated that the invention is intended tocover all modifications and equivalents within the scope of thefollowing claims.

1. A scaffold structure for growing fungi comprising: a. a nutrientsubstrate; b. a porous material defining an intermediate layer whichdoes not readily bind with fungal tissue, wherein the porous material isselected from the group consisting of metal, plastic, and ceramic plate;c. a closed administrable space positioned away from said substrate andsaid porous material; d. a first layer of fungal tissue connecting saidsubstrate to and through said porous material to said administrablespace; e. a successive layer of fungal tissue within said administrablespace; f. a growth field comprising growth field locations such thatgrowth of said first layer of fungal tissue is directed through saidgrowth field locations so as to create said successive layer of fungaltissue; and g. a portion of fungal material delaminated from saidintermediate layer.
 2. The scaffold structure for growing fungi of claim1 wherein the fungal material is at least 50% pure.
 3. The scaffoldstructure for growing fungi of claim 2 wherein the fungal material is100% pure.
 4. The scaffold structure for growing fungi of claim 1wherein the intermediate layer is impermeable with the exception ofporous regions.
 5. The scaffold structure for growing fungi of claim 1wherein the intermediate layer is rigid.
 6. The scaffold structure forgrowing fungi of claim 1 wherein fungal hyphae in the successive layerare fused with at least one additional fungal hyphae.
 7. The scaffoldstructure for growing fungi of claim 1 further comprising a plasticizingagent added to said fungal material.
 8. A scaffold structure for growingfungi comprising: a. a nutrient substrate; b. fungal tissue growing fromsaid nutrient substrate, the fungal tissue comprising fungal hyphae; c.a porous material positioned away from said nutrient substrate, definingan intermediate layer that does not readily bind with said fungaltissue, and through which the fungal tissue extends as a first fungalmaterial layer, wherein the porous material is selected from the groupconsisting of metal, plastic, and ceramic plate; d. an enclosedadministrable space into which said fungal tissue extends as at leastone successive fungal material layer, above which extend fungal hyphaefrom said fungal tissue; and e. a first portion of fungal tissuedelaminated from the intermediate layer, wherein said fungal hyphae aredistorted such that at least one individual hypha comes into contactwith at least one additional fungal hypha.
 9. The scaffold structure ofclaim 8 wherein the fungal tissue is at least 50% pure.
 10. The scaffoldstructure of claim 8 wherein said fungal tissue is joined to the firstfungal material layer and the successive fungal material layer.
 11. Thescaffold structure of claim 8 wherein said hyphae are fused to oneanother.
 12. The scaffold structure of claim 8 wherein said firstportion of fungal tissue delaminated from the intermediate layer is incontact with a second portion of fungal tissue delaminated from theintermediate layer.
 13. The scaffold structure of claim 8 furthercomprising a composite material added to said fungal tissue.
 14. Thescaffold structure of claim 8 wherein said intermediate layer providesuniform initial conditions of growth, thereby achieving uniform growthpattern of the fungal tissue and directing said growth pattern into adefinable plane.
 15. A scaffold structure for growing fungi comprising:a. a nutrient substrate; b. a porous material defining an intermediatelayer selected from the group consisting of metal, plastic, and ceramicplate; c. a closed administrable space positioned away from saidsubstrate and said porous material; d. a first layer of fungal tissueconnecting said substrate to and through said porous material to saidadministrable space; e. a successive layer of fungal tissue within saidadministrable space; a. a growth field comprising growth field locationssuch that growth of said first layer of fungal tissue is directedthrough said growth field locations so as to create said successivelayer of fungal tissue, the successive layer of fungal tissue comprisingfungal hyphae having geometries, wherein said fungal hyphae geometriesare distorted such that at least one individual hypha extending abovethe growth field comes into contact with at least one additional fungalhypha.
 16. The scaffold structure for growing fungi of claim 15 furthercomprising a portion of fungal material delaminated from saidintermediate layer
 17. The scaffold structure for growing fungi of claim15 wherein the intermediate layer is impermeable with the exception ofporous regions.
 18. The scaffold structure for growing fungi of claim 15wherein the intermediate layer is rigid.
 19. The scaffold structure forgrowing fungi of claim 15 wherein fungal hyphae in the successive layerare fused with at least one additional fungal hyphae.
 20. The scaffoldstructure for growing fungi of claim 15 further comprising aplasticizing agent added to said fungal material.